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Göransson, Ulf
Alternative names
Publications (10 of 117) Show all publications
Jacobsson, E., Andersson, H. S., Strand, M., Peigneur, S., Eriksson, C., Lodén, H., . . . Göransson, U. (2018). Peptide ion channel toxins from the bootlace worm, the longest animal on Earth. Scientific Reports, 8, Article ID 4596.
Open this publication in new window or tab >>Peptide ion channel toxins from the bootlace worm, the longest animal on Earth
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2018 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 8, article id 4596Article in journal (Refereed) Published
Abstract [en]

Polypeptides from animal venoms have found important uses as drugs, pharmacological tools, and within biotechnological and agricultural applications. We here report a novel family of cystine knot peptides from nemertean worms, with potent activity on voltage-gated sodium channels. These toxins, named the alpha-nemertides, were discovered in the epidermal mucus of Lineus longissimus, the 'bootlace worm' known as the longest animal on earth. The most abundant peptide, the 31-residue long alpha-1, was isolated, synthesized, and its 3D NMR structure determined. Transcriptome analysis including 17 species revealed eight alpha-nemertides, mainly distributed in the genus Lineus. alpha-1 caused paralysis and death in green crabs (Carcinus maenas) at 1 mu g/kg (similar to 300 pmol/kg). It showed profound effect on invertebrate voltage-gated sodium channels (e.g. Blattella germanica Na(v)1) at low nanomolar concentrations. Strong selectivity for insect over human sodium channels indicates that a-nemertides can be promising candidates for development of bioinsecticidal agents.

Place, publisher, year, edition, pages
NATURE PUBLISHING GROUP, 2018
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-351585 (URN)10.1038/s41598-018-22305-w (DOI)000428029600001 ()29567943 (PubMedID)
Funder
Swedish Research Council, 2014-3327]
Available from: 2018-05-29 Created: 2018-05-29 Last updated: 2018-05-29Bibliographically approved
Strömstedt, A. A., Park, S., Burman, R. & Göransson, U. (2017). Bactericidal activity of cyclotides where phosphatidylethanolamine-lipid selectivity determines antimicrobial spectra. Biochimica et Biophysica Acta - Biomembranes, 1859(10), 1986-2000
Open this publication in new window or tab >>Bactericidal activity of cyclotides where phosphatidylethanolamine-lipid selectivity determines antimicrobial spectra
2017 (English)In: Biochimica et Biophysica Acta - Biomembranes, ISSN 0005-2736, E-ISSN 1879-2642, Vol. 1859, no 10, p. 1986-2000Article in journal (Refereed) Published
Abstract [en]

Cyclotides are a family of plant peptides characterized by a cystine knot embedded in a macrocyclic backbone. They bind to and disrupt phospholipid membranes, which explain their lytic activity on cells. In this study, we expose the full antibacterial potency of cyclotides by avoiding its inhibition by rich growth media assay conditions. For that purpose a two-step microdilution assay protocol was developed, using non-growing conditions during initial peptide incubation. A diverse set of cyclotides was tested for antibacterial and antifungal activity, and the results show that most cyclotides are active under these conditions, especially against Gram-negative bacteria. Activity was observed at sub-micromolar concentrations for three of the cyclotides tested, surpassing that of the control peptides LL-37 and melittin. Noteworthy, two anionic cyclotides were active on Pseudomonas aeruginosa at low micromolar concentrations. Broad-spectrum activity was pronounced among cycloviolacin cyclotides, which included activity on Staphylococcus aureus and Candida albicans. The factors influencing their bactericidal spectrum were revealed by correlating antimicrobial activity with membrane permeabilization on various liposome systems and with the physiochemical properties of the cyclotides. Whereas general electrostatic and hydrophobic parameters are more important for broad-spectrum cyclotides; a phospholipid-specific mechanism of membrane permeabilization, through interaction with phosphatidylethanolamine-lipids, is essential for cyclotides active primarily on Gram-negative bacteria.

Place, publisher, year, edition, pages
ELSEVIER SCIENCE BV, 2017
Keyword
Cyclotide, Antimicrobial peptide, Antibacterial, Membrane permeabilization, Phosphatidylethanolamine-binding, Structure-activity relationship
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-336446 (URN)10.1016/j.bbamem.2017.06.018 (DOI)000411419000024 ()28669767 (PubMedID)
Funder
Swedish Society of Medicine, SLS-254511Swedish Research Council, 2012-5063
Available from: 2017-12-14 Created: 2017-12-14 Last updated: 2017-12-14Bibliographically approved
Hellinger, R., Thell, K., Vasileva, M., Muhammad, T., Gunasekera, S., Kuemmel, D., . . . Gruber, C. W. (2017). Chemical Proteomics for Target Discovery of Head-to-Tail Cyclized Mini-Proteins. Frontiers in Chemistry, 5, Article ID 73.
Open this publication in new window or tab >>Chemical Proteomics for Target Discovery of Head-to-Tail Cyclized Mini-Proteins
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2017 (English)In: Frontiers in Chemistry, E-ISSN 2296-2646, Vol. 5, article id 73Article in journal (Refereed) Published
Abstract [en]

Target deconvolution is one of the most challenging tasks in drug discovery, but a key step in drug development. In contrast to small molecules, there is a lack of validated and robust methodologies for target elucidation of peptides. In particular, it is difficult to apply these methods to cyclic and cysteine-stabilized peptides since they exhibit reduced amenability to chemical modification and affinity capture; however, such ribosomally synthesized and post-translationally modified peptide natural products are rich sources of promising drug candidates. For example, plant-derived circular peptides called cyclotides have recently attracted much attention due to their immunosuppressive effects and oral activity in the treatment of multiple sclerosis in mice, but their molecular target has hitherto not been reported. In this study, a chemical proteomics approach using photo-affinity crosslinking was developed to determine a target for the circular peptide [T20K]kalata B1. Using this prototypic nature-derived peptide enabled the identification of a possible functional modulation of 14-3-3 proteins. This biochemical interaction was validated via competition pull down assays as well as a cellular reporter assay indicating an effect on 14-3-3-dependent transcriptional activity. As proof of concept, the presented approach may be applicable for target elucidation of various cyclic peptides and mini-proteins, in particular cyclotides, which represent a promising class of molecules in drug discovery and development.

Place, publisher, year, edition, pages
FRONTIERS MEDIA SA, 2017
Keyword
cyclotides, cyclic protein, chemical proteomics, peptide-protein interaction, photo-affinity labeling
National Category
Pharmaceutical Sciences
Identifiers
urn:nbn:se:uu:diva-338516 (URN)10.3389/fchem.2017.00073 (DOI)000412726000001 ()29075625 (PubMedID)
Funder
Australian Research Council, FT140100730
Available from: 2018-01-15 Created: 2018-01-15 Last updated: 2018-01-15Bibliographically approved
Park, S., Yoo, K.-O., Marcussen, T., Backlund, A., Jacobsson, E., Rosengren, K. J., . . . Göransson, U. (2017). Cyclotide Evolution: Insights from the Analyses of Their Precursor Sequences, Structures and Distribution in Violets (Viola). Frontiers in Plant Science, 8, Article ID 2058.
Open this publication in new window or tab >>Cyclotide Evolution: Insights from the Analyses of Their Precursor Sequences, Structures and Distribution in Violets (Viola)
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2017 (English)In: Frontiers in Plant Science, ISSN 1664-462X, E-ISSN 1664-462X, Vol. 8, article id 2058Article in journal (Refereed) Published
Abstract [en]

Cyclotides are a family of plant proteins that are characterized by a cyclic backbone and a knotted disulfide topology. Their cyclic cystine knot (CCK) motif makes them exceptionally resistant to thermal, chemical, and enzymatic degradation. By disrupting cell membranes, the cyclotides function as host defense peptides by exhibiting insecticidal, anthelmintic, antifouling, and molluscicidal activities. In this work, we provide the first insight into the evolution of this family of plant proteins by studying the Violaceae, in particular species of the genus Viola. We discovered 157 novel precursor sequences by the transcriptomic analysis of six Viola species: V. albida var. takahashii, V. mandshurica, V. orientalis, V. verecunda, V. acuminata, and V. canadensis. By combining these precursor sequences with the phylogenetic classification of Viola, we infer the distribution of cyclotides across 63% of the species in the genus (i.e., ~380 species). Using full precursor sequences from transcriptomes, we show an evolutionary link to the structural diversity of the cyclotides, and further classify the cyclotides by sequence signatures from the non-cyclotide domain. Also, transcriptomes were compared to cyclotide expression on a peptide level determined using liquid chromatography-mass spectrometry. Furthermore, the novel cyclotides discovered were associated with the emergence of new biological functions.

Keyword
cyclotide evolution, viola phylogeny, sequence signature, cyclotide precursor, neofunctionality, novel cyclotide, precursor domain
National Category
Plant Biotechnology
Identifiers
urn:nbn:se:uu:diva-339782 (URN)10.3389/fpls.2017.02058 (DOI)000418117800001 ()29326730 (PubMedID)
Funder
Swedish Research Council, 2012-5063
Available from: 2018-02-09 Created: 2018-02-09 Last updated: 2018-02-09Bibliographically approved
Malik, S. Z., Linkevicius, M., Göransson, U. & Andersson, D. I. (2017). Resistance to the Cyclotide Cycloviolacin O2 in Salmonella enterica Caused by Different Mutations That Often Confer Cross-Resistance or Collateral Sensitivity to Other Antimicrobial Peptides. Antimicrobial Agents and Chemotherapy, 61(8), Article ID e00684-17.
Open this publication in new window or tab >>Resistance to the Cyclotide Cycloviolacin O2 in Salmonella enterica Caused by Different Mutations That Often Confer Cross-Resistance or Collateral Sensitivity to Other Antimicrobial Peptides
2017 (English)In: Antimicrobial Agents and Chemotherapy, ISSN 0066-4804, E-ISSN 1098-6596, Vol. 61, no 8, article id e00684-17Article in journal (Refereed) Published
Abstract [en]

Antimicrobial peptides (AMPs) are essential components of innate immunity in all living organisms, and these potent broad-spectrum antimicrobials have inspired several antibacterial development programs in the past 2 decades. In this study, the development of resistance to the Gram-negative bacterium-specific peptide cycloviolacin O2 (cyO2), a member of the cyclotide family of plant miniproteins, was characterized in Salmonella enterica serovar Typhimurium LT2. Mutants isolated from serial passaging experiments in increasing concentrations of cyO2 were characterized by whole-genome sequencing. The identified mutations were genetically reconstituted in a wild-type background. The additive effect of mutations was studied in double mutants. Fitness costs, levels of resistance, and cross-resistance to another cyclotide, other peptide and nonpeptide antibiotics, and AMPs were determined. A variety of resistance mutations were identified. Some of these reduced fitness and others had no effect on fitness in vitro, in the absence of cyO2. In mouse competition experiments, four of the cyO2-resistant mutants showed a significant fitness advantage, whereas the effects of the mutations in the others appeared to be neutral. The level of resistance was increased by combining several individual resistance mutations. Several cases of cross-resistance and collateral sensitivity between cyclotides, other AMPs, and antibiotics were identified. These results show that resistance to cyclotides can evolve via several different types of mutations with only minor fitness costs and that these mutations often affect resistance to other AMPs.

Keyword
antimicrobial peptide resistance, cyclotide, cycloviolacin O2, cross-resistance, collateral sensitivity, Salmonella enterica, antimicrobial peptides, fitness, mechanisms of resistance, Salmonella enterica serovar Typhimurium
National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:uu:diva-333077 (URN)10.1128/AAC.00684-17 (DOI)000406259400067 ()
Funder
Swedish Research Council
Available from: 2017-11-06 Created: 2017-11-06 Last updated: 2018-01-13Bibliographically approved
Uddin, S. J., Muhammad, T., Shafiullah, M., Slazak, B., Rouf, R. & Göransson, U. (2017). Single-step purification of cyclotides using affinity chromatography. Biopolymers, 108(3), Article ID e23010.
Open this publication in new window or tab >>Single-step purification of cyclotides using affinity chromatography
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2017 (English)In: Biopolymers, ISSN 0006-3525, E-ISSN 1097-0282, Vol. 108, no 3, article id e23010Article in journal (Refereed) Published
Abstract [en]

Cyclotides are considered promising scaffolds for drug development owing to their inherent host defence activities and highly stable structure, defined by the cyclic cystine knot. These proteins are expressed as complex mixtures in plants. Although several methods have been developed for their isolation and analysis, purification of cyclotides is still a lengthy process. Here, we describe the use of affinity chromatography for the purification of cyclotides using polyclonal IgG antibodies raised in rabbits against cycloviolacin O2 and immobilized on NHS-activated Sepharose columns. Cycloviolacin O2 was used as a model substance to evaluate the chromatographic principle, first as a pure compound and then in combination with other cyclotides, that is, bracelet cyclotide cycloviolacin O19 and Mobius cyclotide kalata B1, and in a plant extract. We demonstrate that single-step purification of cyclotides by affinity chromatography is possible but cross reactivity may occur between homologue cyclotides of the bracelet subfamily.

Keyword
affinity chromatography, cyclotides, cycloviolacin O2, immobilized IgG
National Category
Biochemistry and Molecular Biology Biophysics
Identifiers
urn:nbn:se:uu:diva-334107 (URN)10.1002/bip.23010 (DOI)000407992300005 ()
Funder
Swedish Research Council, 2012-5063
Available from: 2017-11-24 Created: 2017-11-24 Last updated: 2017-11-24Bibliographically approved
Kirkpatrick, C. L., Broberg, C. A., McCool, E. N., Lee, W. J., Chao, A., McConnell, E. W., . . . Hicks, L. M. (2017). The "PepSAVI-MS" Pipeline for Natural Product Bioactive Peptide Discovery. Analytical Chemistry, 89(2), 1194-1201
Open this publication in new window or tab >>The "PepSAVI-MS" Pipeline for Natural Product Bioactive Peptide Discovery
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2017 (English)In: Analytical Chemistry, ISSN 0003-2700, E-ISSN 1520-6882, Vol. 89, no 2, p. 1194-1201Article in journal (Refereed) Published
Abstract [en]

The recent increase in extensively drug-resistant bacterial pathogens and the associated increase of morbidity and mortality demonstrate the immediate need for new antibiotic backbones with novel mechanisms of action. Here, we report the development of the PepSAVI-MS pipeline for bioactive peptide discovery. This highly versatile platform employs mass spectrometry and statistics to identify bioactive peptide targets from complex biological samples. We validate the use of this platform through the successful identification of known bioactive peptides from a botanical species, Viola odorata. Using this pipeline, we have widened the known antimicrobial spectrum for V. odorata cyclotides, including antibacterial activity of cycloviolacin O2 against A. baumannii. We further demonstrate the broad applicability of the platform through the identification of novel anticancer activities for cycloviolacins by their cytotoxicity against ovarian, breast, and prostate cancer cell lines.

National Category
Analytical Chemistry Medicinal Chemistry
Identifiers
urn:nbn:se:uu:diva-316030 (URN)10.1021/acs.analchem.6b03625 (DOI)000392458100025 ()
Available from: 2017-02-24 Created: 2017-02-24 Last updated: 2018-01-13
Gunasekera, S., Fernandes-Cerqueira, C., Eriksson, C., Jakobsson, P.-J. & Göransson, U. (2016). Anti-Citrullinated Peptide Antibody Inhibitors Based On Sunflower Trypsin Inhibitor-1 Scaffold For Potential Anti-Rheumatoid Arthritis Activity. Journal of Peptide Science, 22(S2), S170-S170
Open this publication in new window or tab >>Anti-Citrullinated Peptide Antibody Inhibitors Based On Sunflower Trypsin Inhibitor-1 Scaffold For Potential Anti-Rheumatoid Arthritis Activity
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2016 (English)In: Journal of Peptide Science, ISSN 1075-2617, E-ISSN 1099-1387, Vol. 22, no S2, p. S170-S170Article in journal, Meeting abstract (Other academic) Published
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-331382 (URN)10.1002/psc.2950 (DOI)000398216100317 ()
Available from: 2017-10-17 Created: 2017-10-17 Last updated: 2017-10-17Bibliographically approved
Malik, S. Z., Göransson, U. & Andersson, D. I. (2016). Bacterial Resistance To A Cyclic Antimicrobial Peptide. Journal of Peptide Science, 22(S2), S180-S180
Open this publication in new window or tab >>Bacterial Resistance To A Cyclic Antimicrobial Peptide
2016 (English)In: Journal of Peptide Science, ISSN 1075-2617, E-ISSN 1099-1387, Vol. 22, no S2, p. S180-S180Article in journal, Meeting abstract (Other academic) Published
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-331383 (URN)10.1002/psc.2950 (DOI)000398216100337 ()
Available from: 2017-10-17 Created: 2017-10-17 Last updated: 2017-10-17Bibliographically approved
Gunasekera, S., Fernandes-Cerqueira, C., Eriksson, C., Jakobsson, P. J. & Göransson, U. (2016). Circular disulfide-rich peptide scaffolds as anti-citrullinated peptide antibody inhibitors. Paper presented at 9th Joint Meeting of AFERP, ASP, GA, JSP, PSE and SIF, JUL 24-27, 2016, Copenhagen, DENMARK. Planta Medica, 82
Open this publication in new window or tab >>Circular disulfide-rich peptide scaffolds as anti-citrullinated peptide antibody inhibitors
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2016 (English)In: Planta Medica, ISSN 0032-0943, E-ISSN 1439-0221, Vol. 82Article in journal, Meeting abstract (Other academic) Published
Keyword
anti-citrullinated protein/peptide antibodies (ACPA), cyclic peptide, fibrinogen, NMR, target, sunflower trypsin inhibitor 1 (SFTI-1)
National Category
Pharmacology and Toxicology
Identifiers
urn:nbn:se:uu:diva-346853 (URN)10.1055/s-0036-1596779 (DOI)000411789300603 ()
Conference
9th Joint Meeting of AFERP, ASP, GA, JSP, PSE and SIF, JUL 24-27, 2016, Copenhagen, DENMARK
Available from: 2018-03-27 Created: 2018-03-27 Last updated: 2018-03-27Bibliographically approved
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